Cadherins ; genetics ; Hearing Loss, Sensorineural ; genetics ; pathology ; Humans

OBJECTIVETo study the relationship between E-cadherin gene expression and the methylation status of E-cadherin 5' CpG islands in acute myeloid leukemia (AML).

METHODSReverse transcription-PCR (RT-PCR), flow cytometry and methylation specific PCR were used to analyze the E-cadherin gene and protein expression and its 5' CpG island methylation status respectively in bone marrow cells from 55 AML patients and 7 normal controls.

RESULTSAML cells displayed a significant reduction or lack of E-cadherin gene and protein expression, the positive rates were 23.6% and 18.2% (P < 0.01), respectively. All normal bone marrow cells were E-cadherin positive. Thirty-eight of the 55 patients (69.1%) were E-cadherin 5' CpG island methylated whereas the normal controls were completely unmethylated. Twenty-nine of thirty-one (93.5%) E-cadherin-negative samples showed abnormal hypermethylation of the E-cadherin CpG islands.

CONCLUSIONExpression downregulation and methylation of E-cadherin gene in AML suggest that it might be an important event in AML. E-cadherin methylation was associated with the inhibition of E-cadherin gene and protein expression in AML.

Cadherins ; genetics ; CpG Islands ; genetics ; DNA Methylation ; Gene Expression ; Humans ; Leukemia, Myeloid, Acute ; genetics

Febrile seizures are the most common nervous system disease in childhood, and most children have a good prognosis. However, some epilepsy cases are easily induced by fever and are characterized by "fever sensitivity", and it is difficult to differentiate such cases from febrile seizures. Epilepsy related to fever sensitivity includes hereditary epilepsy with febrile seizures plus, Dravet syndrome, and
Cadherins/genetics* ; Child ; Epilepsy/genetics* ; Epileptic Syndromes ; Humans ; Mutation ; Seizures, Febrile/genetics*

OBJECTIVETo evaluate the role of E-cadherin (E-cad) and CDH1 gene encoding E-cad in the occurrence of sporadic or hereditary gastric cancer.

METHODSNineteen normal gastric mucosal issue specimens, 19 specimens of hereditary gastric cancer (diagnosed according to ICG-HGC criteria), and 19 specimens of sporadic gastric cancer examined for E-cad expression and CDH1 promoter methylation using immunohistochemistry and methylation-specific PCR (MSP).

RESULTSThe protein expression of E-cad were significantly reduced in both of the cancer tissues (P<0.001) compared with that in the normal gastric mucosal tissues, and showed no significant difference between the two cancers (P=0.84). CDH1 promoter hypermethylation was found in 10 out of the 19 hereditary gastric cancer tissues, a rate significantly higher than that in sporadic gastric cancer tissues (3/19, P<0.01).

CONCLUSIONCDH1 promoter hypermethylation is probably an important factor contributing to reduced E-cad expression in sporadic gastric cancer but not in hereditary gastric cancer.

Cadherins ; genetics ; metabolism ; DNA Methylation ; Humans ; Promoter Regions, Genetic ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVE: To analyze the clinical and genetic characteristics of a Chinese pedigree affected with developmental and epileptic encephalopathy 9. METHODS: N048: epilepsy full version gene detection panel-V2 and genome wide copy number variation analysis were carried out on the genomic DNA extracted from the peripheral blood samples. Amniotic fluid was also sampled for single nucleoticle polymorphism array (SNP-array) analysis. RESULTS: Both the mother and her daughter were found to have loss of heterozygosity at Xq21.31q22.1, with which exons of protocadherin 19 (PCDH19) gene was deleted. SNP-array showed the fetus to be a female and had arr[hg19]Xq21.31q22.1 (89 558 626-99 701 006)x1. The mother, daughter and fetus of this family all had developmental and epileptic encephalopathy 9. CONCLUSION Variant of the PCDH19 gene probably underlay the Developmental and epileptic encephalopathy 9 in this pedigree.
Cadherins/genetics* ; China ; DNA Copy Number Variations ; Epilepsy/genetics* ; Epilepsy, Generalized ; Female ; Humans ; Mutation ; Pedigree ; Protocadherins

A large number of putative risk genes for autism spectrum disorder (ASD) have been reported. The functions of most of these susceptibility genes in developing brains remain unknown, and causal relationships between their variation and autism traits have not been established. The aim of this study was to predict putative risk genes at the whole-genome level based on the analysis of gene co-expression with a group of high-confidence ASD risk genes (hcASDs). The results showed that three gene features - gene size, mRNA abundance, and guanine-cytosine content - affect the genome-wide co-expression profiles of hcASDs. To circumvent the interference of these features in gene co-expression analysis, we developed a method to determine whether a gene is significantly co-expressed with hcASDs by statistically comparing the co-expression profile of this gene with hcASDs to that of this gene with permuted gene sets of feature-matched genes. This method is referred to as "matched-gene co-expression analysis" (MGCA). With MGCA, we demonstrated the convergence in developmental expression profiles of hcASDs and improved the efficacy of risk gene prediction. The results of analysis of two recently-reported ASD candidate genes, CDH11 and CDH9, suggested the involvement of CDH11, but not CDH9, in ASD. Consistent with this prediction, behavioral studies showed that Cdh11-null mice, but not Cdh9-null mice, have multiple autism-like behavioral alterations. This study highlights the power of MGCA in revealing ASD-associated genes and the potential role of CDH11 in ASD.
Animals ; Autism Spectrum Disorder/genetics* ; Brain ; Cadherins/genetics* ; Gene Expression ; Mice ; Mice, Knockout

OBJECTIVES: To study the significance of E-cadherin and the association between E-cadherin methylation status and prognosis in children with acute lymphoblastic leukemia (ALL) by examining the mRNA and protein expression of E-cadherin and its gene methylation status in bone marrow mononuclear cells of children with ALL. METHODS: The samples of 5 mL bone marrow blood were collected from 42 children with ALL who were diagnosed for the first time at diagnosis (pre-treatment group) and on day 33 of induction chemotherapy (post-treatment group). RT-qPCR, Western blot, and methylation-specific PCR were used to measure the mRNA and protein expression of E-cadherin and the methylation level of the E-cadherin gene. The changes in each index after induction chemotherapy were compared. RESULTS: The mRNA and protein expression levels of E-cadherin in the post-treatment group were significantly higher than those in the pre-treatment group (P<0.05), while the positive rate of E-cadherin gene methylation in the post-treatment group was significantly lower than that in the pre-treatment group (P<0.05). At the end of the test, the children with negative methylation had significantly higher overall survival rate and event-free survival rate than those with positive methylation (P<0.05). CONCLUSIONS E-cadherin expression is associated with the development of ALL in children, and its decreased expression and increased methylation level may indicate a poor prognosis.
Child ; Humans ; Cadherins/genetics* ; DNA Methylation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics* ; Prognosis ; RNA, Messenger

In order to construct a lentiviral vector carrying human VE-cadherin gene, and to express VE-cadherin in Sup-B15 cells, the VE-cadherin gene was amplified by RT-PCR from the human placenta, and then cloned into pCR-Blunt vector. The VE-cadherin DNA fragment was subcloned into pLB vector to generate a lentiviral vector pLB-VEC. Recombinant lentivirus was generated by co-transfection of three-plasmids into 293FT packing cells using lipofectamine 2000. The Sup-B15 cells were transfected by the lentivirus. The post-transfected Sup-B15 cells were observed by microscopy and flow cytometry. Western blot was used to determine the expression of VE-cadherin. The results showed that the VE-cadherin DNA fragment was amplified from human placenta and was cloned into pCR-Blunt vector, the recombinant lentiviral vector pLB-VEC was successfully constructed. High titer lentivirus was prepared by 3-plasmid packing system, and transfected into Sup-B15 cells in vitro effectively. The obviously morphological changes occurred in transfected cells, the expression of VE-cadherin protein could be detected in Sup-B15 cells via flow cytometry and Western blot. It is concluded that the lentiviral vector pLB-VEC carrying human VE-cadherin gene is successfully constructed; VE-cadherin gene is expressed in Sup-B15 cells via lentiviral vector transfection, which provides an optional tool for further study on the mechanism of VE-cadherin controlling leukemia development.
Antigens, CD ; genetics ; Cadherins ; genetics ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Recombinant Fusion Proteins ; genetics ; Transfection

To examine the expression of forkhead transcription factor O4 (FOXO4) in prostate cancer and to explore its effect on prostate cancer cell invasion.
 Methods: Immunohistochemistry was used to detect the expression of FOXO4 in prostate hyperplasia tissues and prostate cancer tissues. Western blot was used to detect the expression of FOXO4 in prostate hyperplasia cell line BPH-1 and prostate cancer cell lines: PC-3 and DU145. PC-3 cells with high relative expression of FOXO4 were transfected with FOXO4 siRNA and scramble siRNA; DU145 cells with low expression of FOXO4 were transfected with FOXO4 plasmid and blank vector. Matrigel Transwell assay was used to detect the invasive ability of transfected cells. The expression of endothelial-mesenchymal transition (EMT)-related proteins E-cadherin, N-cadherin, and vimentin in the transfected cells was detected by Western blot.
 Results: The expression of FOXO4 in prostate cancer cells and tissues was significantly lower than that in the prostate hyperplasia cells and tissues (both P<0.05). In the prostate cancer tissues, the expression of FOXO4 in cancer tissues with prostate cancer specific antigen (PSA) value <4 was significantly higher than that in the tissues with 4≤PSA≤10 and PSA>10 (all P<0.05). The expression of FOXO4 in cancer tissues with Gleason score <8 was significantly higher than that in the cancer tissues with Gleason ≥8 (P<0.05). The expression of FOXO4 in clinical stage T1-T2 prostate cancer tissues was higher than that in the clinical stage T3-T4 prostate cancer tissues (P<0.05). The expression of FOXO4 in prostate cancer tissues without lymph node metastasis was significantly higher than that in the prostate cancer tissues with lymph node metastasis (P<0.05). Down-regulation of FOXO4 in PC-3 cells could significantly promote the EMT and invasion, with the decreased expression of E-cadherin and the increased expression of N-cadherin and vimentin (all P<0.05); Up-regulation of FOXO4 in DU145 cells could inhibit the EMT and invasion of cells, with the increased expression of E-cadherin and the decreased expression of N-cadherin and vimentin (all P<0.05).
 Conclusion: FOXO4 is involved in prostate cancer progression, and it can inhibit prostate cancer cell invasion by regulating EMT of prostate cancer cells.
Cadherins ; genetics ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Neoplasm Invasiveness ; genetics ; Prostatic Neoplasms ; genetics ; Transcription Factors ; genetics

OBJECTIVETo investigate the protein expression, methylation promoter, somatic and germ-line mutations of E-cadherin gene (CDH1) in hereditary gastric cancer in China and to investigate its possible roles.

METHODSEight probands diagnosed with ICG-HGC criterion were enrolled in our database from June 1994 to October 2007. Tumor tissues were detected for CDH1 expression by using immunohistochemistry (IHC) methods. CDH1 DNA sequencing was performed for all its 16 exons both in tumor and normal tissues of the same patients to detect somatic and germ-line mutations. Methylation promoter study was performed by using specific primers and polymerase chain reaction (PCR) methods.

RESULTSIHC analysis confirmed that the CDH1 expression was negative in 7 probands and downregulated in the other on proband. Six mutations in five probands were found with DNA sequencing: two silent mutations and four missense mutations. All six mutations were absent in normal tissues, thereby excluded its presence in germ-line cells. Both DNA missense mutations and gene silencing through promoter methylation was found in 4 probands. Two probands has only promoter methylation and one proband had only silent mutation. No DNA missense mutations or promoter methylation was found in one proband.

CONCLUSIONSCDH1 gene germ-line mutations are relatively rare in hereditary gastric cancer in China, and whereas CDH1 somatic mutations and promoter methylation synergistically induce CDH1 downregulation in these patients.

Cadherins ; genetics ; DNA Methylation ; DNA Mutational Analysis ; Germ-Line Mutation ; Humans ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics